SUMMARYWe have studied the synthesis of virus RNA in human embryo lung cells infected with rhinovirus type 2. The three species of RNA in extracts of infected ceils are, in order of decreasing electrophoretic mobility, single-stranded RNA, replicative form and replicative intermediate. The kinetics of synthesis of these RNA species were investigated. The electrophoresis of mixtures of single-stranded RNA synthesized early and late in infection showed no differences in size. This observation was confirmed with a bovine enterovirus. The native form of poliovirus type I singlestranded RNA migrated faster on polyacrylamide electrophoresis than rhinovirus single-stranded RNA.
Ribonuclease treatment of rhinovirus-infected human embryo lung cells after cell disruption reveals that double stranded RNA is present in the preparation before nucleic acids are extracted with phenol. This shows that the hydrogen bonding between complementary molecules of viral RNA which occurs in infected cells is not a result of the extraction of RNA with phenol.
The kinetics of appearance of an RNA-dependent RNA polymerase activity obtained from human embryo lung cells infected with rhinovirus type 2 have been followed by analysis of the RNA synthesized by the polymerase preparation in vitro. Little single-stranded RNA was synthesized and the proportion of replicative intermediate to replicative form was over threefold greater than obtained in vivo. The polymerase activity in vivo declined in the presence of cycloheximide showing that continued protein synthesis was necessary to maintain RNA replication.
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